Murine Kidney Transplant Technique

Summary

The goal of this manuscript is to describe the steps required to perform a kidney transplant in a mouse, paying particular attention to the details of the arterial anastomosis.

Abstract

The first mouse kidney transplant technique was published in 1973¹ by the Russell laboratory. Although it took some years for other labs to become proficient in and utilize this technique, it is now widely used by many laboratories around the world. A significant refinement to the original technique using the donor aorta to form the arterial anastomosis instead of the renal artery was developed and reported in 1993 by Kalina and Mottram ² with a further advancement coming from the same laboratory in 1999 ³. While one can become proficient in this model, a search of the literature reveals that many labs still experience a high proportion of graft loss due to arterial thrombosis. We describe here a technique that was devised in our laboratory that vastly reduces the arterial thrombus reported by others ^4,5. This is achieved by forming a heel-and-toe cuff of the donor infra-renal aorta that facilitates a larger anastomosis and straighter blood flow into the kidney.

Introduction

Since 1973 the kidney transplant model in mice has been a valuable research tool, but technical issues have hampered its widespread use. Over the years several papers have been published detailing improvements/refinements to this procedure. As a model of primarily vascularized solid organ transplantation this procedure is probably second only to the heterotopic heart transplant model which was also devised by the Russell laboratory in 1973 ⁶. Both models lend themselves to research into allogeneic rejection responses, the development of delayed graft function and ischemia reperfusion injury.

One of the most common issues to be re....

Protocol

All mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and were housed under pathogen free conditions at the University of Colorado Denver, Barbara Davis Center Animal Facility according to NIH Guidelines and with approval of the University of Colorado Denver IACUC.

1. Donor Kidney Harvest

1. Sterilize all instruments, wear sterile gloves throughout the procedure and maintain a sterile field. Perform all surgeries with the use of an operating microscope.
2. Surgically remove mouse donor kidneys from pentobarbital-anesthetized (60mg/kg IP) donors. Ascertain depth of anesthesia by toe-pinch, and observe the respi....

Results

This surgical technique allows for either simple graft survival/rejection studies, or quite complex experimental protocols. In the figures below we demonstrate the advantages of using this improved arterial anastomosis technique. Using this technique we have significantly reduced the incidence of arterial thrombosis from 35% to 0% thus increasing productivity. We have used this technique for over one year with the same 0% thrombosis result maintained. Figure 1 describes the method for the formation of th.......

Discussion

Mastering this transplant technique is difficult, but once accomplished it is a very powerful research tool. The patient surgeon/researcher will be rewarded by attention to detail and consistency of technique, which is the key to mastering any surgical procedure, even more so in small animal models. The technical difficulties of mastering the mouse kidney transplant are many folds, and it is highly probable that experience in other small animal transplant models must be gained before tackling this procedure.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Instrument	Roboz #	Fine Science Tools #	Arosurgical #
Straight micro-dissecting forcep #5	RS-5015	11295-51
Curved micro-dissecting forcep #7	RS-5047	11297-00
Curved serrated forcep	RS-5137	11052-10
Vannas micro-dissecting scissors, short	RS-5610		09.140.08
Micro-dissecting scissors, straight, sharp, long		11.602.11
Micro spring handle needle holder			11.549.15
Straight mosquito forcep		91308-12
Micro-dissecting scissors, straight, blunt	RS-5962	14078-10
Micro-dissecting scissors, curved, blunt	RS-5981	14079-10
Micro retractor	RS-6540
Instrument tray, 10” x 6 ½” x ¾”	RT-1350S
Silk suture, 5/0, 22.5m spool		18020-50
Suture
10/0 nylon			T4A10Q07
5/0 silk			E19A05N
Gloves		Drapes
Biogel from Medex Supply		Precept, #64-9012-9
Syringes		Cotton applicators
B-D 1cc insulin, #329424		Fisher-brand, #23-400-100
Povidone-Iodine swabs
PDI, #B40600
4/0 Cotton ties
Domestic cotton autoclaved with instruments